BACKGROUND AND OBJECTIVES: Previously, various methodologies were used to enumerate the endothelial progenitor cells (EPCs). We now know that these methodologies enumerate at least three different EPC subsets: circulating angiogenic cells (CACs), colony-forming unit endothelial cells (CFU-ECs), and endothelial colony-forming cells (ECFCs). It is not clear whether there is a correlation between changes in the number of these subsets. The aim of the current study is to find an answer to this question. MATERIALS AND METHODS: The number of all EPC subsets was quantified in the peripheral blood of nine pregnant women in their first and third trimesters of pregnancy. We enumerated 14 cell populations by quantitative flow-cytometry using various combinations of the markers, CD34, CD133, CD309, and CD45, to cover most of the reported phenotypes of CACs and ECFCs. Culturing technique was used to enumerate the CFU-ECs. Changes in the number of cells were calculated by subtracting the number of cells in the first trimester peripheral blood from the number of cells in the third trimester peripheral blood, and correlations between these changes were analyzed. RESULTS: The number of CFU-ECs did not correlate with the number of ECFCs and CACs. Also, CACs and ECFCs showed independent behaviors. However, the number of CACs showed a strong correlation with the number of CD133+CD309+ cells (p=0.001) and a moderate correlation with the number of CD34+CD309+ cells (p=0.042). Also, the number of ECFCs was correlated with the number of CD309+CD45- cells (p=0.029) and CD34+CD45- cells (p=0.03). CONCLUSION: Our study showed that the three commonly used methods for quantifying EPC subsets represent different cells with independent behaviors. Also, any study that measured the number of EPCs using the flow cytometry method with a marker combination that lacks CD309 may be inaccurate.
Endothelial Cells ; Endothelium ; Female ; Flow Cytometry ; Humans ; Phenotype ; Pregnancy ; Pregnancy Trimester, First ; Pregnancy Trimester, Third ; Pregnant Women ; Stem Cells*

OBJECTIVE: In vitro fertilization (IVF) is a well-known method for the treatment of infertility. The present study aimed to compare the differences between infertile women with successful and unsuccessful IVF outcomes regarding the expression of T helper (Th) cell transcription factors and a group of related cytokines before and after exposure to their husbands' seminal plasma. METHODS: This study was performed on 19 couples with unexplained infertility undergoing IVF treatment. Among the studied group, nine and 10 couples had successful and unsuccessful IVF outcomes, respectively. This study was carried out using real-time polymerase chain reaction. RESULTS: Before seminal plasma exposure, the expression levels of T-bet (p < 0.007), interferon-γ (p=0.013), and tumor necrosis factor (TNF)-α (p=0.017) were higher in the infertile women with IVF failure than in those with successful IVF outcomes, while those of GATA3 (p < 0.001), Foxp3 (p=0.001), and interleukin (IL)-35 (p < 0.003) were lower. After seminal exposure, the expression of T-bet (p=0.02), Rorc (p < 0.001), TNF-α (p=0.001), Foxp3 (p=0.02), and interferon-γ (p=0.001) increased in the unsuccessful IVF group, while the expression of Foxp3 (p=0.02), Rorc (p < 0.001), IL-23 (p=0.04), IL-17 (p=0.02), IL-6 (p < 0.001), transforming growth factor-β (p=0.01), and IL-35 (p < 0.001) increased in the successful IVF group. CONCLUSION: In summary, IVF failure was associated with imbalanced Th1/Th2/Th17/Treg responses. Moreover, our results show that seminal plasma might have a positive effect on IVF outcomes via changes in peripheral blood T cell subsets.
Cytokines* ; Family Characteristics ; Female ; Fertilization in Vitro* ; Humans ; In Vitro Techniques* ; Infertility ; Interleukin-17 ; Interleukin-23 ; Interleukin-6 ; Interleukins ; Methods ; Real-Time Polymerase Chain Reaction ; Semen* ; T-Lymphocyte Subsets ; T-Lymphocytes, Helper-Inducer* ; Transcription Factors ; Tumor Necrosis Factor-alpha

OBJECTIVE: In order to increase the number of mature oocytes usable for intracytoplasmic sperm injection (ICSI), we aimed to investigate the effect of co-culturing granulosa cells (GCs) on human oocyte maturation in vitro, the fertilization rate, and embryo development. METHODS: A total of 133 immature oocytes were retrieved and were randomly divided into two groups; oocytes that were cultured with GCs (group A) and oocytes that were cultured without GCs (group B). After in vitro maturation, only oocytes that displayed metaphase II (MII) underwent the ICSI procedure. The maturation and fertilization rates were analyzed, as well as the frequency of embryo development. RESULTS: The mean age of the patients, their basal levels of follicle-stimulating hormone, and the number of oocytes recovered from the patients were all comparable between the two study groups. The number of oocytes that reached MII (mature oocytes) was 59 out of 70 (84.28%) in group A, compared to 41 out of 63 (65.07%) in group B (p=0.011). No significant difference between fertilization rates was found between the two study groups (p=0.702). The embryo development rate was higher in group A (33/59, 75%) than in group B (12/41, 42.85%; p=0.006). The proportion of highest-quality embryos and the blastocyst formation rate were significantly lower in group B than in group A (p=0.003 and p<0.001, respectively). CONCLUSION: The findings of the current study demonstrate that culturing immature human oocytes with GCs prior to ICSI improves the maturation rate and the likelihood of embryo development.
Blastocyst ; Coculture Techniques* ; Embryonic Development* ; Embryonic Structures* ; Female ; Fertilization ; Follicle Stimulating Hormone ; Granulosa Cells* ; Humans* ; In Vitro Oocyte Maturation Techniques ; Metaphase ; Oocytes* ; Pregnancy ; Sperm Injections, Intracytoplasmic